Imaging Energy Status in Live Cells With a Fluorescent Biosensor of the Intracellular ATP-to-ADP Ratio

Nat Commun. 2013;4:2550. doi: 10.1038/ncomms3550.

Abstract

The ATP:ADP ratio is a critical parameter of cellular energy status that regulates many metabolic activities. Here we report an optimized genetically encoded fluorescent biosensor, PercevalHR, that senses the ATP:ADP ratio. PercevalHR is tuned to the range of intracellular ATP:ADP expected in mammalian cells, and it can be used with one- or two-photon microscopy in live samples. We use PercevalHR to visualize activity-dependent changes in ATP:ADP when neurons are exposed to multiple stimuli, demonstrating that it is a sensitive reporter of physiological changes in energy consumption and production. We also use PercevalHR to visualize intracellular ATP:ADP while simultaneously recording currents from ATP-sensitive potassium (KATP) channels in single cells, showing that PercevalHR enables the study of coordinated variation in ATP:ADP and KATP channel open probability in intact cells. With its ability to monitor changes in cellular energetics within seconds, PercevalHR should be a versatile tool for metabolic research.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adenosine Diphosphate / analysis*
  • Adenosine Diphosphate / biosynthesis
  • Adenosine Triphosphate / analysis*
  • Adenosine Triphosphate / biosynthesis
  • Animals
  • Archaeal Proteins / genetics
  • Archaeal Proteins / metabolism
  • Astrocytes / metabolism*
  • Astrocytes / ultrastructure
  • Biosensing Techniques*
  • Cell Line
  • Embryo, Mammalian
  • Energy Metabolism / physiology
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Green Fluorescent Proteins / analysis
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • KATP Channels / genetics
  • KATP Channels / metabolism
  • Mice
  • Microscopy, Fluorescence, Multiphoton
  • Molecular Imaging
  • Neurons / metabolism*
  • Neurons / ultrastructure
  • Primary Cell Culture
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Single-Cell Analysis

Substances

  • Archaeal Proteins
  • GlnK1 protein, Methanosarcina mazei
  • KATP Channels
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • Adenosine Diphosphate
  • Adenosine Triphosphate